Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h
The introduction of direct detectors and the automation of data collection in cryo-EM have led to a surge in data, creating new opportunities for advancing computational processing. In particular, on-the-fly workflows that connect data collection with three-dimensional reconstruction would be valuab...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
International Union of Crystallography
2017-11-01
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| Series: | IUCrJ |
| Subjects: | |
| Online Access: | http://scripts.iucr.org/cgi-bin/paper?S205225251701226X |
| _version_ | 1818083777511948288 |
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| author | Björn O. Forsberg Shintaro Aibara Dari Kimanius Bijoya Paul Erik Lindahl Alexey Amunts |
| author_facet | Björn O. Forsberg Shintaro Aibara Dari Kimanius Bijoya Paul Erik Lindahl Alexey Amunts |
| author_sort | Björn O. Forsberg |
| collection | DOAJ |
| description | The introduction of direct detectors and the automation of data collection in cryo-EM have led to a surge in data, creating new opportunities for advancing computational processing. In particular, on-the-fly workflows that connect data collection with three-dimensional reconstruction would be valuable for more efficient use of cryo-EM and its application as a sample-screening tool. Here, accelerated on-the-fly analysis is reported with optimized organization of the data-processing tools, image acquisition and particle alignment that make it possible to reconstruct the three-dimensional density of the 70S chlororibosome to 3.2 Å resolution within 24 h of tissue harvesting. It is also shown that it is possible to achieve even faster processing at comparable quality by imposing some limits to data use, as illustrated by a 3.7 Å resolution map that was obtained in only 80 min on a desktop computer. These on-the-fly methods can be employed as an assessment of data quality from small samples and extended to high-throughput approaches. |
| first_indexed | 2024-12-10T19:43:23Z |
| format | Article |
| id | doaj.art-9ac30ee046bb42b0bb374c25ac6bd725 |
| institution | Directory Open Access Journal |
| issn | 2052-2525 |
| language | English |
| last_indexed | 2024-12-10T19:43:23Z |
| publishDate | 2017-11-01 |
| publisher | International Union of Crystallography |
| record_format | Article |
| series | IUCrJ |
| spelling | doaj.art-9ac30ee046bb42b0bb374c25ac6bd7252022-12-22T01:35:57ZengInternational Union of CrystallographyIUCrJ2052-25252017-11-014672372710.1107/S205225251701226Xkf5004Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 hBjörn O. Forsberg0Shintaro Aibara1Dari Kimanius2Bijoya Paul3Erik Lindahl4Alexey Amunts5Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenScience for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenScience for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenScience for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenScience for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenScience for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 171 65 Solna, SwedenThe introduction of direct detectors and the automation of data collection in cryo-EM have led to a surge in data, creating new opportunities for advancing computational processing. In particular, on-the-fly workflows that connect data collection with three-dimensional reconstruction would be valuable for more efficient use of cryo-EM and its application as a sample-screening tool. Here, accelerated on-the-fly analysis is reported with optimized organization of the data-processing tools, image acquisition and particle alignment that make it possible to reconstruct the three-dimensional density of the 70S chlororibosome to 3.2 Å resolution within 24 h of tissue harvesting. It is also shown that it is possible to achieve even faster processing at comparable quality by imposing some limits to data use, as illustrated by a 3.7 Å resolution map that was obtained in only 80 min on a desktop computer. These on-the-fly methods can be employed as an assessment of data quality from small samples and extended to high-throughput approaches.http://scripts.iucr.org/cgi-bin/paper?S205225251701226Xcryo-EMimage processingchlororibosome |
| spellingShingle | Björn O. Forsberg Shintaro Aibara Dari Kimanius Bijoya Paul Erik Lindahl Alexey Amunts Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h IUCrJ cryo-EM image processing chlororibosome |
| title | Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h |
| title_full | Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h |
| title_fullStr | Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h |
| title_full_unstemmed | Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h |
| title_short | Cryo-EM reconstruction of the chlororibosome to 3.2 Å resolution within 24 h |
| title_sort | cryo em reconstruction of the chlororibosome to 3 2 a resolution within 24 h |
| topic | cryo-EM image processing chlororibosome |
| url | http://scripts.iucr.org/cgi-bin/paper?S205225251701226X |
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